Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1) and U-87 MG (Lane 2). The blots were probed with Anti-SOD1 Mouse Monoclonal Antibody (Product # MA1-105, 2 µg/ml) and detected using Goat anti-Mouse IgG (H and L) Secondary Antibody, DyLight 800 4X PEG (Product # SA5-35521) at dilutions 1:10,000 (Fig. 1) and 1:30,000 (Fig. 2). A 18 kDa band corresponding to SOD1 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody after blocking with 5 % skimmed milk. Fluorescent detection was performed using the Odyssey® Fc imaging system (Li-cor Biosciences).
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Conjugate||DyLight 800 4X PEG|
|Purification||Antigen affinity chromatography|
|Storage buffer||proprietary buffer|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:100 - 1:2000|
|Immunohistochemistry (IHC)||1:100 - 1:2000|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:10,000-1:30,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Product SA5-35521 reacts with the heavy chains of Mouse IgG and with the light chains common to most Mouse immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.
To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol without and store at -20°C. Note: not compatible with sodium azide. Mole Dye/Mole Protein Ratio is lot-dependent.
DyLight 800-4xPEG has an excitation (nm) of 783 nm (Amax) and an emission (nm) of 797 nm (Emax) in PBS. The emission spectra for DyLight™ 800-4xPEG conjugate match the principal output wavelengths of most common fluorescence instrumentation.
Thermo Scientific Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for Mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.